Targeted glycoprotein enrichment and identification in stromal cell secretomes using azido sugar metabolic labeling.

ABSTRACT:

Purpose

Effectively identifying the proteins present in the cellular secretome is complicated due to the presence of cellular protein leakage and serum protein supplements in culture media. A metabolic labeling and click chemistry capture method is described that facilitates the detection of lower abundance glycoproteins in the secretome, even in the presence of serum.

Experimental design

Two stromal cell lines were incubated with tetraacetylated sugar-azide analogs for 48 h in serum-free and low-serum conditions. Sugar-azide labeled glycoproteins were covalently linked to alkyne-beads, followed by on-bead trypsin digestion and MS/MS. The resulting glycoproteins were compared between media conditions, cell lines, and azide-sugar labels.

Results

Alkyne-bead capture of

SUBMITTER: Roper SM 

PROVIDER: S-EPMC4331031 | biostudies-literature | 2013 Jun

REPOSITORIES: biostudies-literature